Passive shedding of erythrocyte antigens induced by membrane rigidification |
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Authors: | Claue P Muller Meir Shinitzky |
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Institution: | Department of Membrane Research, The Weizmann Institute of Science, Rehovot 76100, Israel |
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Abstract: | Rigidification of the cell membrane lipid bilayer can lead to an increase in the degree of exposure of membrane proteins to either side of the membrane. It is shown in this study that excess increase of the membrane lipid microviscosity (‘hyper-rigidification’) in intact human erythrocytes can cause the release of Rh0(D) and A blood group antigens from the cell surface which can then be collected from the supernatant by affinity chromatography. The most efficient antigen shedding was achieved upon incorporation of cholesteryl hemisuccinate (CHS) (incubation for 2 h at 37 °C in a mixture of 200 μg/ml CHS, 3.5% polyvinylpyrrolidone 1% bovine serum albumin, 0.5% glucose in phosphate-buffered saline) followed by application of hydrostatic pressure (1 500 atm, 5 min) which increases the lipid microviscosity by about 2-fold. This technique can be of general application for isolation of membrane proteins without disruption of the cells or the use of detergents. |
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Keywords: | To whom offprint requests should be addressed |
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